title: NMDA receptor subunit expression in the supraoptic nucleus of adult rats: dominance of NR2B and NR2D.
abstract: The supraoptic nucleus (SON) of the hypothalamus contains magnocellular neurosecretory neurons (MNC) which synthesize and release the peptide hormones vasopressin and oxytocin. Glutamate is a prominent excitatory neurotransmitter in the SON and regulates MNC excitability. NMDA receptors (NMDAR), a type of ionotropic glutamate receptor, mediate synaptic plasticity of MNCs and are necessary for characteristic burst firing patterns which serve to maximize hormone release. NMDARs are di- or tri-heteromeric complexes of NR1 and NR2 subunits. Receptor properties depend on NR2 subunit composition and variable splicing of NR1. We investigated the expression profile of NR1 and NR2 subunits in the SON at the mRNA and protein levels plus protein expression of NR1 splice variants in control and salt-loaded adult rats. There was robust mRNA expression of all subunits, with NR2D levels being the highest. At the protein level, NR1, NR2B, and NR2D were robustly expressed, while NR2A was weakly expressed. NR2C protein was not detected with either of the two antibodies tested. All four NR1 splice variant cassettes (N1, C1, C2, C2') were detected in the SON, although NR1 N1 expression was too low for accurate analysis. Three days of salt-loading did not alter mRNA, protein, or splice variant expression of NMDAR subunits in the SON. Robust NR2D protein expression has not been previously shown in MNCs and is uncommon in the adult brain. Although the functional significance of this unusual expression profile is unknown, it may contribute to important physiological characteristics of SON neurons, such as burst firing and resistance to excitotoxicity.
authors.affiliation: University of Colorado, School of Medicine, Department of Physiology and Biophysics, RC-1 North Tower, P18-7127, Mail Stop 8307, Aurora, CO 80045, USA. Faye.Doherty@world.oberlin.edu
authors.forename: Faye C
authors.initials: FC
authors.lastname: Doherty
authors.forename: Celia D
authors.initials: CD
authors.lastname: Sladek
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003681:Dehydration / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D008297:Male; D020033:Protein Isoforms / Q000096:biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013495:Supraoptic Nucleus / Q000378:metabolism*
chemical_list: C409466:NR1 NMDA receptor; C121001:NR2B NMDA receptor; C464436:NR2D NMDA receptor; D020033:Protein Isoforms; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate